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Appl. Environ. Microbiol. doi:10.1128/AEM.01158-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Detection and identification of fungi intimately associated with the brown seaweed Fucus seratus

Institute of Phytopathology and Applied Zoology, University of Giessen, D-35392 Giessen, GERMANY; Deptartment of Botany and Plant Pathology, 2082 Cordley Hall, Oregon State University, Corvallis, OR 97331, USA; Institute of Marine Sciences, University of North Carolina, Morehead City, N. C. 28557, USA; School of Biological Sciences, University of Portsmouth, Portsmouth PO1 2DY, UK

^* To whom correspondence should be addressed. Email: Alga.Zuccaro{at}agrar.uni-giessen.de.

	Abstract

The filamentous fungi associated with healthy and decaying Fucus serratus thalli were studied over a one-year period using isolation methods and molecular techniques such as 28S-rDNA PCR-DGGE, phylogenetic and real-time PCR analyses. The predominant DGGE bands from healthy algal thalli belonged to the Lindra, Lulworthia, Engyodontium, Sigmoidea/Corollospora-complex and Emericellopsis/Acremonium-like ribotypes. In the culture-based analysis Sigmoidea marina isolates had the highest incidence of recovery. In general, the environmental sequences retrieved could be matched unambiguously to isolates recovered from the seaweed, with the exceptions of the Emericellopsis/Acremonium-like ribotype, which showed 99% homology with the sequences from four different isolates, including that for Acremonium fuci. To estimate the extent of colonization of A. fuci we used a TaqMan real time quantitative PCR assay for intron 3 of the Beta-tubulin gene; the probe for which proved to be species-specific even when used in amplifications with high background concentrations of other eukaryotic DNAs. A. fuci sequence was detected from both, healthy and decaying thalli with stronger signal in the latter. Additional sequence types, representing members from the Dothideomycetes, were recovered from the decaying thalli DNA, which suggested that a change in fungal community structure had occurred. Phylogenetic analysis of these environmental sequences with those from isolates and type species indicated that they were novel within the Dothideomycetes.